Pregna-4, 6-diene-and 19-norpregna-4, 6-diene-3beta, 16, 17alpha-triol steroids, derivatives thereof, and processes for their preparation



ABSTRACT OF THE DISCLOSURE Novel pregna-4,6-dieneand 19-norpregna-4,6-diene steroids of the pregnane series and derivatives thereof containing substitution at the C3, C-6, C-16, and C-17a positions which compounds are progestational and antiinfiammatory agents and processes for the preparation of such compounds.

This invention relates to novel cyclopentanopolyhydrophenanthrene derivatives and to processes for the preparation thereof.

More particularly, this invention relates to novel A pregnadieneand 19-nor-A -pregnadiene-3B,17a-diol derivatives represented by the general formula:

In the above formula R represents hydrogen or methyl, R represents hydrogen or an acyl group, R represents hydrogen, a-methyl, fl-methyl, u-hydroxyl, or uacyloxy, CR and R taken together represent the groupmg:

wherein R and R represent hydrogen, an alkyl (including cycloalkyl) group or an aryl (including aralkyl and alkaryl) group containing up to 8 carbon atoms, inclusive, and X represents hydrogen, methyl, fluoro, chloro or bromo.

The acyloxy and acyl groups referred to herein are preferably derived from hydrocarbon carboxylic acids containing less than 12 carbon atoms. These acids can be saturated or unsaturated (including aromatic), and can have straight, branched, cyclic or cyclic-aliphatic chains. In addition, they can be substituted with one or more functional groups, such as hydroxyl groups, alkoxy groups containing up to 6 carbon atoms, acyloxy groups containing up to 12 carbon atoms, nitro groups, amino groups or halogen atoms. Typical ester groups are the acetate, proprionate, enanthate, benzoate, trimethylacetate, tbutylacetate, phenoxyacetate, cyclopentylpropionate, aminoacetate, and fi-chloropropionate groups.

The novel A -pregnadieneand 19-nor-A -pregnadiene-3fl,17a-diol derivatives represented by Formula A hereinabove are progestational agents which also possess anti-androgenic activity and are also useful in the treatment of menstrual disorders and in fertility control.

United States Patent 0 3,386,994 Patented June 4, 1968 These novel A -pregnadieneand 19-nor-A -pregnadiene-3B,17a-diol derivatives can be prepared by processes which can be illustrated schematically as follows:

""351; a Q R U X X l o: no-

---orr "-OR5 A MR2 MR2 .l .r

RO- R50- IV 111 Ir X on, on, l ---on -----0 R3 --orr x c i 0 \R4 R R v VI l CH3 $113 1 "-03 ---orr om l --Ol[ R l R l R5O R10- VIII v11 (3H3 (1:0 mom I --on R In these formulas R, R R R, R and X have the same meanings as set forth hereinabove for Formula A, and R represents an acyl group.

The starting materials represented by Formula I hereinabove are prepared in the manner described in US. Patent No. 3,138,589 to Ringold et al.

In carrying out the above-illustrated processes, the starting material (I), e.g., 6-chloro-A -pregnadien-17aol-3,20-dione 17-acetate (I; R=methyl, R =acetyl, R =hydrogen, X=chloro), is dissolved in an inert organic solvent, e.g., a lower alkanol such as methanol or ethanol, an ether such as dioxane or tetrahydrofuran, or the like, and reacted under substantially anhydrous conditions with from about 0.5 mol to about mols, and preferably from about 1 to about 4 mols, per mol of steroid starting material, of sodium borohydride, lithium borohydride, or the like, at a temperature ranging from about 0 C. to about 60 C., and preferably at room temperature for from about 10 minutes to about 12 hours, to give the corresponding 35-01 (II), e.g., 6-chloro-A pregnadiene-3B,17a-diol-20-one 17-acetate (II; R=methyl, R =acetyl, R =hydrogen, X=chloro).

The thus-obtained 313-01 can be acylated, using an anhydride or chloride of an organic acid, e.g., one derived from an acid of the type referred to hereinabove, in pyridine or the like at room temperature, thus giving the corresponding 3,17-diacylate (III). For example, by reacting 6-chloro-A -pregnadiene-3,3,l7a-diol-20-one 17- acetate with acetic anhydride in pyridine at room temperature for 20 hours, the corresponding 3,17-diacetate (III; R=methyl, R =acetyl, R =hydrogen, X=chloro) is obtained.

The l7ot-acyloxy group in the 35-01 (II) can also be hydrolyzed, e.g., by boiling with a 15% solution of sodium hydroxide, potassium hydroxide, or the like, in a lower alkanol under an inert nitrogen atmosphere for from about 1 hour to about 3 hours or longer, to produce the corresponding 35, 170t-dIOI (IV), e.g., 6-chloro-A pregnadiene-S/B, 17a-di0l-20-0116 (IV; R=methyl, R and R'-=hydrogen, X=chloro).

By reacting the thus-obtained 3,3, l7a-diol (IV) with an acid anhydride or chloride in pyridine or the like in the manner described hereinabove, the corresponding 3- monoacylate (IV), e.g., 6-chloro-A -pregnadiene-3B, 17a-diol-20-one 3-acetate (IV; R=methyl, R =acetyl, R =hydrogen, X=chloro), is obtained.

When a 16-unsubstituted-3B, 17oc-di0l (IV), e.g., 6- chloro-A -pregnadiene-Iifi, 17u-diol-20-one, is incubated in known manner with an oxygenating microorganism such as Streptomyces roseochromogenus or the like, the corresponding 3,8, 16oz, 17zx-tli0l (V), e.g., 6-Chloro- A -pregnadiene-3B, 16m, 17rx-triol-20-one (V; R=methyl, X=chloro), is produced.

The thus-prepared 3,8, 160:, 17outri0l can be reacted with an aldehyde or ketone in the presence of a catalytic amount of a strong acid, such as perchloric acid, ptoluenesulfonic acid, or the like, in the manner described in US. Patent No. 3,048,581 to Fried, thus giving the corresponding 16a, 17a-cyclic acetal or ketal (VI), e.g., 6 chloro-l6a, 17ct-isopropylidenedioxy-M -pregnadien- 3fi-ol-20-one (VI; R, R and R =methyl, R =hydrogen, X=chloro).

Esterification of the free 3,8-hydroxyl group in the resulting 16cc, 17u-cyclic acetal or ketal with an acid anhydride or chloride in pyridine or the like in the manner described hereinabove gives the corresponding 3-acylate (VI), e.g., 6-chloro-16a, 17a-is'opropylidenedioxy-A pregnadien-3B-ol-20-one 3-acetate (VI; R, R and R methyl, R =acetyl, X=chloro).

Hydrolysis of the 160:, 17a-CyCliC acetal or ketal grouping in the thus-prepared 3-acylate, using an aqueous 60- 90% solution of formic acid at a temperature of from about 50 C. to about 110 C. for from about 1 minute to about 1 hour, gives the corresponding 3,3-acyloxy-16a, 170L-di01 (VII), e.g., 6-chloro-A -pregnadiene-3B, 16:1, l7a-triol-20-one 3-acetate (VII; R=methyl, R =acetyl, X=chloro).

By reacting either the 3,8-acyloxy- 160a, i70t-dlo1 (VII) or the 35, 16a, 17oc-t1l0l (V) with a mixture of an organic acid and the corresponding acid anhydride at room temperature in the presence of a catalytic amount of a strong acid, such as p-toluenesulfonic acid or the like, the corresponding 3, 16, 17-triacylate (VIII) is obtained. For example, by reacting 6-chloro-A -pregnadiene-3,8, 16oz, 17a-triol-20-one or its 3-acet ate with a mixture of acetic acid and acetic anhydride in the presence of a catalytic amount of p-toluenesulfonic acid, the corresponding 3, 16, l7-triacetate (VIII; R=methyl, R =acety1, X=chloro) is obtained.

The 3/3- and 16a-acyloxy groups in the thus-obtained 3, 16, 17-triacy1ate can be selectively hydrolyzed, using a dilute aqueous solution of a strong base in a lower alkanol, preferably a 15% aqueous methanolic solution of sodium hydroxide or potassium hydroxide, at room temperature for from about 10 minutes to about 10 hours, thus giving the corresponding 3,3, 16a, 17a-triol 17-acylate (IX), e.g., 6-chloroA -pregnadiene-3,3, 160:, 17atriol-20-one 17-acetate (IX; R=methyl, R =hydrogen, R =acetyl, X=chloro).

The free 3,3-hydroxyl group in the resulting 3,9, 160:, 17a-triol 17-acylate can then be selectively reacylated by reacting said 17-acylate with an acid anhydride, on a mol-for-mol basis, in pyridine or the like at a temperature of about 20 C. or below, thus giving the corresponding 3,8, 160:, l7a-triol, 3, 17-diacylate (IX). For example, by reacting 6-chloro-A -pregnadiene-3,8, 16a, 17oa-tIlOl-20- one l7-acetate with acetic anhydride, on a mol-for-mol basis, in pyridine at 20 C., the corresponding 3, 17-diacetate (IX; R=methyl, R and R =acetyl, X=chloro) is obtained.

In those cases wherein any of the esterification reactions described hereinabove are carried out on steroid starting materials wherein one hydroxyl group has pre-- viously been esterified, esterifying agents different from those previously employed can be used, if desired, to give mixed esters. For example, by esterifying 6-chloro-A pregnadiene-3/3, 16a, 17a-triol-20-one l7-acetate with propionic anhydride in the manner just described, the corresponding 3 propionate-l7-acetate (IX; R=methyl, R =propionyl, R =acetyl, X=chloro) is obtained.

In order that those skilled in the art can more fully understand the present invention, the following examples are set forth. These examples are given solely for the purpose of illustrating the invention, and should not be considered as expressing limitations unless so set forth in the appended claims.

PREPARATION A A mixture of 5 grams of l6a-methyl-l7m-acetoxy progesterone, grams of chloranil and 200 cc. of t-butanol was refluxed for 10 hours. Following this reaction period the reaction mixture was cooled and the excess of chloranil filtered off and washed with a large volume of ethyl acetate. These washings were then added to the filtrate, and the combined solution was washed with a cold, aqueous 10% sodium hydroxide solution until the washings were colorless. The organic layer (chiefly ethyl acetate, containing the product) was then dried over anhydrous sodium sulfate and evaporated to dryness. Recrystallization of the resulting residue from methylene dichloride/diethyl ether gave 16a-methyl-A -pregnadien-17aol-3,20-dione 17-acetate.

By repeating this procedure in every detail but one, namely, replacing l6a-methyl-l7a-acetoxy progesterone with 16tx-methyl-19-nor-17a-acetoxy progesterone, 16,6- rnethyl-17ot-acetoxy progesterone and 16,6-methyl-19-norl7a-acetoxy progesterone, the corresponding 6-dehydro steroids, i.e., 16a-methyl-19-nor-A -pregnadien 17a ol- 3,20-dione 17-acetate, 16,3-methyl-A -pregnadien-17a-ol- 3,20-dione 17-acetate and 1fifi-mcthyl-l9-nor-A -pregnadien-l7u-ol-3,20-dione l7-acetate, respectively were ob tained.

5 PREPARATION B A solution of 4 grams of loot-methyl-l9-nor-A -pregnadien-l7a-ol-3,20-dione 17-acetate in 100 cc. of chloroform was cooled to C., then admixed with 4 molar equivalents of monoperphthalic acid dissolved in diethyl ether. The resulting reaction mixture was held at room temperature for 20 hours, following which it was diluted with water. Next, the organic layer was separated, washed with an aqueous sodium bicarbonate solution and then with water until neutral, then dried over anhydrous sodium sulfate and finally evaporated to dryness. Recrystallization of the dry residue from acetone/hexane gave 6a,7ot-oxido-l6ut-methyl-l9-nor-A -pregnen-17ot ol 3,20- dione 17-acetate.

One-gram of 6a,7ot-oxido-16a-methyl-19-nor-A -pregnenl7a-ol-3,20-dione l7-acetate was suspended in 35 cc. of glacial acetic acid. A slow current of anhydrous hydrogen chloride gas was passed through the suspension, and was continued for hours. The reaction mixture was then concentrated to about /3 its original volume by distillation under reduced pressure at 35 C., then poured into 7 ice water. The thus-formed precipitate was collected by filtration, washed with water until neutral, and then dried. Recrystallization from methylene dichloride-hexane gave 6-chloro-l6ot-methyl-l9-nor-A pregnadien 170a ol- 3,20-clione 17-acetate.

Similarly, by replacing anhydrous hydrogen chloride gas with anhydrous hydrogen bromide, there is obtained 6-bromo-16ot-methyl-l9-nor-A -pregnadien-l704-01 3,20- dione-17-acetate PREPARATION C In accordance with the method described in Preparation B, 16B-methyl-l9-nor-A -pregnadien-17a ol 3,20- dione-17-acetate, 16fl-methyl-A -pregnadien-17a-ol-3,20- dione-17-acetate and lout-methyl-a z -pregnadien-17a ol- 3,20-dione were converted into the corresponding 6-chloro-16,8-methyl-l9-nor-A prcgnadien 17oz ol 3,20- dione-17-acetate,6-chloro-16B-rnethyl A pregnadien- 17ot-ol-3,20-dione-l7-acetate, 6-chloro-16u methyl A pregnadien-17a-ol-3,20-dione, 6-bro-mo-l6fl methyl 19- nor A pregnadien 17a ol 3,20 dione 17 acetate, 6-bromo-16fl methyl A pregnadien 17cc ol- 3,20-dione-l7-acetate, and 6-bromo-l6a-methyl-A -pregnadien-l7u-ol-3,20-dione 17-acetate.

PREPARATION D A solution of 1 gram of ootflu-oxido-l6a-methyl-l9- nor-M-pregnen-l7ot-ol-3,20-dione 17-acetate in a mixed solvent consisting of 25 cc. of benzene and 25 cc. of diethyl ether was admixed with 5 cc. of freshly distilled boron trifluoride etherate. The resulting reaction mixture was allowed to stand at room temperature overnight, following which it was diluted with water. The organic layer was separated, washed with water, dried over anhydrous sodium sulfate and evaporated to dryness under vacuum, thus giving 6fi-fluoro-l6ot-methyl-l9-nor A pregnene- 7a,17a-dlOl-3,20-dl0n6 17-acetate.

This 6fi-fluoro-7w-hydroxy intermediate was suspended in 35 cc. of glacial acetic acid, and a slow current of anhydrous hydrogen chloride gas was passed through this suspension for 4 hours. Following this reaction period the product was worked up in the manner described in Preparation C hereinabove, thus giving 6-fluoro-l6a-methyl-l9- norA -pregnadien-l7ot-ol-3,20-dionc 17-acetate.

By repeating this entire procedure in every detail but one, namely, using 6a,7a-oxido-lfia-methyl-A -pregnenl7a-ol-3,20-dione 17-acetate, 6a,7ot-oxido-l6fi-methyl-A pregnen-l7a-ol-3,20-dione l7-acetate and 6a,7otOXidO- l6,8-methyl-l9-nor-A -pregnen-17a 01 3,20 dione l7- acetate, was the' steroid starting materials, 6-1'iuoro-l6amcthyl-a -pregnadien-l7a-ol-3,20-dione l7 acetate, 6- fluoro-16,8-methyl-A -pregnadien-Howl-3,20 dione l7- acetate and 6-fluoro-l6,6methyl-19-nor-A pregnadienl7a-ol-3,20-dione l7acetate, respectively, were obtained.

6 PREPARATION E A suspension of 5 g. of 16ot-methyl-6ot-chloro-A -pregnen-l7et-ol-3,20-dione acetate in 37.5 cc. of anhydrous peroxide-free dioxane was treated with 6 cc. of freshly distilled ethyl orthoformate and 4 g. of p-toluenesulfonic acid. The mixture was stirred at room temperature for 15 minutes and the resulting solution let stand for 30 minutes further. 4 cc. of pyridine were added and then water. The formed precipitate was collected by filtration, water washed and air dried. There were thus obtained 3-ethoxy-l6a-methyl-6-chloro-A -pregnadien 170a ol- 3,20-dione acetate which was used for the next step without further purification.

A solution of 5 g. of the foregoing enol ether in 20 cc. of tetrahydrofuran was cooled to 0 C. and there were added 1.05 molar equivalents of 2,3-dichloro-5,6-dicyanol,4benzoquinone and 100 mg. of p-toluenesulfonic acid. The resulting mixture was further stirred at 0 C. for 30 minutes. The precipitated hydroquinone was filtered off and 100 cc. of methylene chloride was added to the filtrate.

The organic solution was washed with 5% aqueous sodium hydroxide solution until the washings were colorless, then with water to neutrality, dried over anhydrous sodium sulfate and evaporated to dryness. Crystallization from acetone-hexane gave l6ot-methyl-6-chloro-A pregnadien-17ot-ol-3,20-dione acetate which was identical with the compound described in Preparation C.

PREPARATION F In accordance with the method described in the precedmg preparation, the compounds listed below under I were converted into the corresponding 6-dehydro derivatives II:

I II

6,lfia-dimethyLN-pregnen-Ua-ol- 3,20'dione propionate.

6a,1(SB-dimethyl-A -pregnen-l7aol-3,20-dione.

To a solution of l g. of the acetate of 6-chloro-6- dehydro-lh-hydroxy progesterone in 50 cc. of ethanol there was added mg. of sodium borohy-dride, and the reaction mixture was kept at room temperature for 40 minutes. There were then cautiously added 5 cc. of acetone and 0.2 cc. of concentrated hydrochloric acid, and the mixture diluted with water. The product was then extracted with ether and the organic extract washed with 5% sodium carbonate solution and water to neutral, dried over anhydrous sodium sulfate and evaporated to dryness. The residue was crystallized from acetone-hexane to produce 840 mg. of 6-chloro-A -pregnadiene-3fi, l7a-diol- 20-one 17-acetate, M.P. 193l95 C.; [ab 63 (CHCI This procedure was then repeated in every detail but one, namely, 6-chloro-A -pregnadien-l7a-ol-3,20-dione l7-acetate was replaced by A -pregnadien-l7ot-ol-3,20-dione 17-acetate,

' 6-fluoro-A -pregnadien-17ot-ol-3,20-dione 17-acetate,

6-chloro-16a-methyl-A -pregnadien-17a-ol-3,20-dione 17-acetate,

6-chloro-1 6rx-n1ethyl-A -pregnadien-17a-ol-3 ,ZO-dione 17-va1erate,

6-bromo-16rx-methyl-A -pregnadien-17a-ol-3,20-dione 17-acetate,

6-fluoro-16fl-methyl-A -pregnadien-17a-ol-3,20-dione 17 -acetate,

6-chloro-16fi-methyl-A -pregnadien-l7a-ol-3 ,20-dione 17-acetate,

6-bromo-1 6,8-methy1-A -pregnadiem 17 oc-O1-3,20-di0n6 l7-acetate,

19-nor-A -pregnadienl7rx-0l-3 ,20-di-one 17-acetate,

6-fiuoro-19-nor-A -pregnadienl7ec-ol-3 ,ZO-dione 17-acetate,

6-chloro-l9-nor-A -pregnadien-17rx-ol-3 ,20-dione 17-acetate,

6-chloro- 19-nor-A -pregnadien- 17oc-01-3,20-di0116 l7-caproate,

6-bromo 19-nor-A -pregnadien- 17ac-0l-3 ,20 dione 17-acetate,

16a-me'thyll9-nor-A -pregnadien-l7a-ol-3 ,ZO-dione 17-acetate,

16 fi-methyll 9-nor-A pre gnadienl 7u-ol-3,20-di-one 17-acetate,

6-fiu-oro-16a-rnet-hy1-19-nor-A -pregnadien-17a-ol-3 ,20-

dione l7-acetate,

6-chloro-16a-methyl-19-nor-A -pregnadien-17a-ol-3,20-

dione 17 -acetate,

6-br0mo- 1 6ot-methyl- 19-nor-A -pregnadien- 1704- 01-3,

20-dione 17-acetate,

6-fiuoro-l 6fl-methyl-19-nor-A -pregnadien-17or-ol-3 ZO-dione 17 -acetate,

6-chlorol 6 8-methyll9-nor-A -pregnadienl7a-o1-3 20-dione 17-acetate, and

6 -bromo- 1 6fl-methyl-19-nor-A -pre gnadien- 17oc-0l-3 20-dion 17-acetate, respectively.

In each case, the corresponding 35-01, namely,

A regnadiene-3/3, 17u-diol-20-one 17-acetate,

6-fluoro-A -pregnadiene-3B, 17a-dio1-20-one 17-acetate,

6-ch'loro-A -pregnadiene-3fl, 17a-diol-20-0ne 17-propionate,

6-chl0ro-A -pregnadiene-Sfi, 17oc-diOl-20-One 17-butyrate,

6-chloro-A -pregnadiene-3B, 17w-diol-20-one l7-cyclopentylpropionate,

6-ch1oro-A -pregnadiene-3p, 17u-diol-20-one 17- caproate,

6-brorno-A -pregnadiene-3B, 17a-di0l-20-0I1e 17-acetate,

16a-methyl-A -pregnadiene-3,B,17u-diol-2O-one 17- acetate,

16b-rnethyl-A -pregnadiene-35, 17a-diol-20-one 17- acetate,

6-fiuoro-16a-methyl-A -pregnadiene-3B, 17a-di0l-20-0ne 17-acetate,

6-chl-oro-16a-methyl-M -pregnadiene-iifi, 17a-diol-20-one 17-acetate,

6-chloro-16or-methy1-A -pregnadiene-3 ,8, l7a-diol-20-one 17-valerate,

6-bron1o-16or-methyl-A pregnadiene-3p, 17or-diol-20-one 17-acetate,

6-fluoro-16,8-methyl-A -pregnadiene-Sfi, 17a-diol-20-one 17-acetate,

6-chloro-16fi-methyl-A -pregna-diene-3fi, 17or-dio1-20-one 17-acetate,

6-bromo-16fl-methyl-A -pregnadiene-3,8, 17a-dio1-20-one 17-acetate,

19-nor-A -pregnadiene-3f3, 17oc-diO1-20-One l7-acetate,

6-fluoro-19-nor-A -pregnadiene-3fi, 170c-dlOl-20-One 17-acetate,

6-chloro19-nor-A -pregnadiene-3fi,17a-dio1-20-one 17-acetate,

6-chloro-19-nor-A -prcgnadiene-SB, 17oc-dlUl20-Ol16 17-caproate,

A mixture of 750 mg. of 6-chloro-A -pregnadiene-Iifl, 17or-diol-20-one l7-acetate, 3 cc. of pyridine and 3 cc. of acetic anhydride was kept at room temperature for 4 hours. It was then diluted with water and the formed precipitate collected by filtration, washed with water and air dried. Crystallization from acetone-ether gave the pure diacetate of 6-chloro-A -pregnadiene-3B,l7a-diol-20-one, M.P. l98201 C.;[rx] -87 (CHCl By repeating this procedure using the remaining Bfl-ols produced in Example I, the corresponding 3,8-acetates thereof are formed.

Example III By repeating the method of Example II except that the acetic anhydride was substituted by propionic anhydride, butyric anhydride, caproic anhydride and enanthic anhydride, there were correspondingly obtained the 3,8-propionates, 3fl-butyrates, 3B-caproates and the 3B-enanthates.

Example IV To a solution of 500 mg. of the 17-acetate of 6-chloro- A -pregnadiene-3B,17a-diol-2O-one in 25 cc. of ethanol there was added 250 mg. of sodium hydroxide, and the reaction mixture was refluxed for 1 hour. It was then diluted with water and the product extracted with ethyl acetate, the organic extract was washed with water to neutral, dried over anhydrous sodium sulfate and evaporated to dryness, thus producing 6-chloro-A -pregnadiene-3B,17a-diol-20-one.

By repeating this procedure in every detail but one, namely, using the remaining 3fl-ol-17a-acylates produced in Example I as the starting material, the corresponding 3,6,17fl-diols were obtained.

Example V 6 chloro-A -pregnadiene-3B,17or-diol-20-one was reacted with acetic anhydride in pyridine in the manner described in Example II hereinabove, thus giving 6-chloro- A -pregnadiene-iifi,l7u-diol-20-one 3-acetate, M.P. 177 C.

By repeating this procedure using the remaining free 35,17a-di0ls prepared as described in Example IV hereinabove as the steroid starting materials, the corresponding 3-acetates were obtained.

Similarly, by replacing acetic anhydride with propionic, butyric, caproic and enanthic anhydrides, and using each of the free 313,17a-di0ls prepared as described in Example IV hereinabove as the steroid starting materials, the corresponding 3-propionates, 3-butyrates, 3-caproates and 3- cnanthates were obtained.

Example VI A culture of Streptonzyces roseoclu'omogenlls, ATCC No. 3347, was prepared in an inclined agar medium containing 1% by weight of glucose and 1% by weight of yeast extract, each based on the total weight of the medium. One cc. portions of a suspension of this culture were then used to innoculat each of a series of 250 cc. Erlenmeyer fiasks which each contained 50 cc. of a sterilized aqueous medium containing 2% by weight of peptone and by weight of corn syrup, each based on the total weight of the sterilized aqueous medium. The resulting mixtures were then incubated, with shaking and under aeration, at 28 C., for 48 hours, thus giving vegetating growing cultures of the microorganism.

Ten mg. of 6-chloro-A -pregnadiene-3[3,17,8-diol-20- one were added to each flask, and the resulting mixtures were incubated, with stirring and under aeration, at 28 C. for 72 hours, following which the mixtures were combined and extracted several times with methylene dichloride. The thus-obtained extract was washed with water,, dried over anhydrous sodium sulfate, filtered, and the filtrate evaporated to dryness under reduced pressure. Chromatography of the resulting residue on silica gel gave 6chloro-A -pregnadiene-3B,16a,17 ix-triol-20-one.

By repeating this procedure in every detail but one, namely, using the remaining 16-unsubstituted-3B,17adiols prepared as described in Example IV hereinabove as the steroid starting materials, the corresponding 36, 16a,17u-triols namely, A -pregnadiene-3/Ll6a,17atriol- 20-one, 6-fluoro-A -pregnadiene-3B,16a,17a-triol-20-one, 6 bromo-A -pregnadiene-3[3,16a,17a-triol-20-one, 19- nor-A -pregnadiene-3fi,16a,17ot-triol-20-one, 6-fiuoro-19- nor-A -pregnadiene-w,16a,17a-triol-20-one, 6-chloro-19- nor-A -pregnadiene-3/8,16a,17a-tri0l-20-one, and 6-bromo 19-nor-A -pregnadiene-3,8,16a,17u-triol-20-one, respectively, were obtained.

Example VII A mixture of 1 gram of 6-chloro-A pregnadiene-3 6, 16a,17a-tIiOl-20-On6, 50 cc. of acetone and 0.5 cc. of an aqueous 72% solution of perchloric acid was allowed to stand at room temperature, with stirring, for 1 hour. Following this reaction period the reaction mixture was washed with an aqueous sodium bicarbonate solution and then with water until neutral, then steam distilled and the product extracted with methylene dichloride. The resulting extract was then dried over anhydrous sodium sulfate and evaporated to dryness. Crystallization of the thus-obtained residue from acetone-hexane gave 6-chloro- 16oz, 17u-isopropylidenedioxy-A -pregnadien-3B-ol-2O-one.

This procedure was then repeated in every detail but one, namely, the remaining 3/3, 160a, 17a-triols prepared as described in Example VI hereinabove were used as the steroid starting materials. In each case, the corresponding 16a, l7a-acetonide, namely, 16oz, l7a-isopropylidenedioxy- A -pregnadien 3fl-ol-2O-one, 6-fluoro-16a, 17OC-ISOPIOPY1- idenedioxy A pregnadien-3fl-ol-20-one, 6-bromo-16ot, 17a-isopropylidenedioxy .A pregnadien-3/i-ol-20-one, 160:, 17a isopropylidenedioxy-l9-nor-A -pregnadien-3B- ol-20-one, 6-fluoro 16a, 17a-isopropylidenedioxy-l9-nor- A -pregnadien-3,8-ol-2O-one, 6 chloro 16a, l7a-is0' propylidenedioxy-lQ-nor A pregnadien-3fi-ol-20-one, and 6-bromo-16a, l7u-isopropylidenedioxy 19-nor-A pregnadien-3fi-ol-20-one, respectively, was obtained.

Similarly by replacing acetone with methyl ethyl ketone, cyclohexanone and acetophenone, and using each of the 3,8, 16m, 17a-triols just mentioned, the corresponding 16oz, 17a-(2-butylidenedioxy), 16a, l7ot-cyclohexylidenedioxy and 16a, 17a-(2'-phenylethylidenedioxy) derivatives were obtained.

Example VIII 6-chloro-16a, 17ot-isopropylidenedioxy-A -pregnadien- 3}3-0l-20-one was reacted with acetic anhydride in pyridine in the manner described in Example II hereinabove, thus giving 6-chloro-16a, 17u-isopropylidenedioxy-A pregnadien-3B-ol-20-one 3-acetate.

By repeating this procedure using the remaining 16a, 17a-isopropylidenedioxy steroids prepared as described in Example VII hereinabove, the corresponding B-acetates were obtained.

Example IX One gram of 6-Ch10l'O-160t, 17a-isopropylidenedioxy- A -pregnadien-3fl-ol-20-one 3-acetate was admixed with 20 cc. of 60% formic acid, and the resulting reaction mixture was heated on a steam bath for 1 hour. Following this reaction period the reaction mixture was cooled to room temperature and diluted with water. The resulting precipitate was collected by filtration, washed with water and dried. Recrystallization from acetone-hexane gave 6- chloro-A -pregnadiene-3B, 16m, 170: triol 20 one 3 acetate.

By repeating this procedure using the remaining l7a-isopropylidendioxy 3-acetates prepared as described in Example VIII hereinabove as the steroid starting materials, the corresponding free 16a, t-dlOlS, namely, A -pregnadiene-3fl, 16a, l7ot-triol-20-one 3-acetate, 6- fiuoro-Ae -pregnadiene-3B, 16a, l7a-triol-20-one 3-acetate, 6-bromo-A -pregnadiene-3fi, 16a, l7a-triol-20-one 3- acetate, 19-nor-A -pregnadiene-3fl, 16a, l7a-triol-20-one 3-acetate, 6-fiuoro-19-nor-A -pregnadiene-3,8, 16cc, 17cctriol-20-one 3-acetate, 6-chloro-19-nor-A -pregnadiene- 3,8, 16a, l7a-triol-20-one Z-acetate, and 6-bromo-19-nor- A -pregnadiene-3fi, 16a, l7ot-triol-20-one 3-acetate, respectively, were obtained.

Example X A mixture of 1 gram of 6-chloro-A -pregnadiene-3fl, 160:, l7a-triol-20-one 3-acetate, 1 gram of p-toluenesulionic acid monohydrate, 0 cc. of acetic acid and 25 cc. of acetic anhydride was allowed to stand for 24 hours at room temperature. Following this reaction period the reaction mixture was poured into water and stirred until the excess acetic anhydride present had hydrolyzed, and this solution was then extracted with methylene dichloride. The thus-obtained extract was then washed with water, dried over anhydrous sodium sulfate and evaporated to dryness. Crystallization of the resulting residue from acetone-diethyl ether gave 6-chloro A -pregnadiene-3fl, 16oz, 17a-triol-20-one 3,16,17-triacetate.

By repeating this procedure using the remaining 3,3, 160:, 17a-triol 3-acetates prepared as described in Example [X hereinabove as the steroid starting materials, the corresponding 3,16,17-triacetates were obtained.

Similarly, by repeating this procedure using the free 318, 16cc, 17a-triols prepared as described in Example VI hereinabove as the steroid starting materials, and replacing the mixture of acetic anhydride and acetic acid with mixtures of propionic anhydride and propionic acid, butyric anhydride and butyric acid, and caproic anhydride and caproic acid, the corresponding 3,16,17 -tripropionates, 3,16,17-tributyrates and 3,16,17-tricaproates were obtained.

Example XI A- solution of 1 gram of 6-chloro-A -pregnadiene-BB, 16a, l7ot-triol-20-ome 3,16,17-tria-cetate in 25 cc. of metha- 1101 was admixed with a solution of 0.6 grams of potassium hydroxide in 5 cc. of water, and the resulting reaction mixture was allowed to stand at room temperature for 3 hours. Following this reaction period the reaction mixture was poured into ice water and the resulting precipitate Was collected by filtration, washed with water until neutral and dried. Recrystallization from methylene dichloridediethyl ether gave 6-chloro-A -pregnadiene-3B, 16a, 17a triol 20-one 17-acetate.

By repeating this procedure using the remaining 3,16,

17-triacetates prepared as described in Example X hereinabove as the steroid starting materials, the corresponding l7-monoacetates, namely, A -pregnadiene-3fi, 16a, 17atriol-20-one l7-acetate 6-fluoro-A pregnadiene-3fi, 16a, 17oc-triol-20-0ne 17-acetate, 6-bromoaA -pregnadiene-3fl, 16oz, 17oc-tfiOl-20-O1'l6 17-acetate, 19-nor-A -pregnadiene- 313, 16a, l7a-triol-20-one, l7-acetate, 6-fiuoro-19-nor-A 1 l pregnadiene-3B, 16cc, 17u-triol-20-one l7-acetate, 6-chloro- 19-nor-A -pregnadiene-3B, 16a, 17a-triol-20-one l7-acetate, and 6 bromo-A -pregnadiene-3B, 16oz, 17ot-triol-20- one 17-acetate, respectively, were obtained.

Example XII A solution of 1 gram of 6 chloro A pregnadiene- 3,8,l6oc,l7oc triol 20 one 17 acetate in 50 ml. of pyridine was admixed with 1 molar equivalent of acetic anhydride, and the resulting reaction mixture was held at 20 C. for 6 hours. Following this reaction period the reaction mixture was poured into ice water and the resulting precipitate was collected by filtration, washed with water and dried. Chromatography of the dry precipitate on 200 times its weigh-t of silica gel, using ethyl acetatehexane (20:80, respectively, by volume) as the eluant, gave 6 chloro A pregnadiene 3fi,l6oc,l7oc triol- 20 one 3,17 diacetate.

By repeating this procedure using the remaining 17- monoacetate prepared as described in Example XI hereinabove as the steroid starting materials, the corresponding 3,17 diacetates were obtained.

Similarly, by replacing acetic anhydride with propionic, butyric and caproic anhydrides, and using each of the 17- monoacetates prepared as described in Example XI hereinabove as the steroid starting materials, the corresponding 3 propionate 17 acetates, 3 butyrate 17 acetates and 3 caproate 17 acetates were obtained.

Example XIII Example XIV By following the method of Example I, the compounds listed below under I were converted into the products set forth under II:

I II

6-rnethyl-A -pregnadien17a-ol- 3,20-(1ione caproate. 16a,17a-isoproylidenedioxy-A pregnadiene-3,20-dione. 6,16a-dirnethyl-A --pregnadien 17aol-3,20-dione. 6-chl0r0 16a,17a-isopropylidenedixy-A -pregnadicne-3,20-di0ue.

one.

6,16a-dimethyl-A -prcgnadien-Hw 6,16a-rlimethyl-A -pregnadieneol-3,20-di0ne propionate. 3fl,17a-diol-20-one 17-propionate.

A -pregnadione-lfia,17a-diol-3,20- A -pregnadiene-3;3,16a,17a-triol (lione diacetate. 20-0nc 17-acetate.

6chlor0-A -pregnadiene-l611,1701- 6 ehl0ro-A pregnadieueBfi, 16a,

diol-3,20-dione diacetatc. 17a-trioL20-onc 17-aeetate.

Example XV By following the esterification method of Example II, 6 methyl A pregnadiene 3fi,l7oz diol 20 one 17 caproate and l6oz,l7oc isopropylidenedioxy A pregnadien 3p ol 20 one were converted respectively into: 6 methyl A pregnadiene 3B,l7oc diol 20- one 3 acetate 17 caproate and 16a,17a isopropylidenedioxy A pregnadien 3B ol 2O one acetate.

Example XVI In accordance with the method described in Example XIII, the compounds mentioned below under I were 12 esterified with the indicated carboxylic acid anhydrides, to produce the corresponding diesters listed under II:

I Anhydride II lfia-methyl-fi-lluoro-N Propionic. 1(lot-moth,vl-fi-r'luoro-A pregnadiene-BB,l7adiolpregnadiene-3B,Not-diol- 20-one 17-acetate. 20-one 3-propionate 17- acetate.

Oaproie 16ct-methyl-6-ehl0r0-A pregnadiene-ZlfiJh-diol- 20-0110 3-caproate 17- acetate.

nadicne-3B,l7a-diol20- one dipropionate.

6,16a-dimeth yl-A -pregnadiellO-BB,l7a-(1l0l-20Dll0 17-aeetate.

A --prcgnadiene-3B,17a- C yclopentyl- A -plcgfitltliGllG-IIBJ7adiol-ZO-one l7-acetate. propionic. diol-20-one S-cyclopentylpropionate 17-acetate. 6-methyl-A -pregnadiene- Caproic 6-methyl-A -pregnadienc- 3fi,17adiol-20-onc 17- 313,17a-d10l-20-0I16 3,17- caproate. dicaproate.

Example XVII Example XVIII In accordance with the method of Example II, 6,160;- dimethyl A pregnadiene 313,170 diol 20 one, 6,165 dimethyl A pregnadiene 36,170: diol 20- one, 16oz methyl A pregnadiene 35,1741 diol 20- 'one, A pregnadiene 3fl,16oc,17a triol 2O one 17- acetate and 6 chloro A pregnadiene 3 3,160t,l7ottriol 2O one 17 acetate were converted respectively into: 6,16ot dimethyl A pregnadiene 318,17a diol- 20 one 3 acetate, 6,165 dimethyl A pregnadiene- 3,B,17u diol 20 one 3 acetate, 16a methyl A pregnadiene 36,17 diol 20 one 3 acetate, A pregnadiene 3,8,16a,17a triol 20 one triacetate and 6 chloro A pregnadiene 3/3,16ot,l7oc triol 20- one triacetate.

Example XIX By the following saponification method of Example IV, A -pregnadiene-3fi,17a-diol-2O-one 17-acetate, 6-fluoro- A -pregnadiene-3fi,17u-diol-20-one 17-acetate, 6-methyl- A -pregnadiene-3fi,17a-diol-2O-one 17-caproate and 6, 16a-dimethyl-A -pregnadiene-3,8,17a-diol-2O-one 17-propionate were converted into the corresponding free compounds.

Example XX To a solution of 1 g. of 6-chloro-A -pregnadiene-3fi, 17ot-diol-20-one in 20 cc. of anhydrous benzene there were added 0.2 g. of p-toluenesulfonic acid and 2 cc. of acetic anhydride and the mixture was allowed to stand for 24 hours at room temperature, poured into ice and water, and the resulting mixture stirred to efiect hydrolysis of the excess anhydride. The benzene layer was separated and washed with 10% sodium carbonate solution and water. Drying, evaporation and crystallization of the residue from ether-hexane produced the diacetate of 6-chloro- A -pregnadiene-3B,17a-diOl-2O-one, identical to that obtained in Example II.

In a similar manner but using propionic, caproic and cyclopentylpropionic anhydrides as esterifying agents there were produced the dipropionate, dicaproate and dicyclopentylpropionate of 6 chloro A -pregnadiene-3fl,17ot- (liol-20-one.

It will be obvious to those skilled in the art that other changes and variations can be made in carrying out the present invention without depatring from the spirit and scope thereof as defined in the appended claims.

wherein R is selected from the group consisting of hydrogen and methyl, R is selected from the group consisting of hydrogen and a hydrocarbon carboxylic acyl group containing less than 12 carbon atoms, R is selected from the group consisting of (Jr-hydrocarbon carboxylic acyloxy and a-hydroxy, OR and R taken together represent the grouping wherein R and R are selected from the group consisting of hydrogen, an alkyl group and an aryl group containing up to 8 carbon atoms, inclusive, and X is selected from the group consisting of hydrogen, methyl, fiuoro, chloro and bromo.

2. Compounds according to claim 1 wherein OR and R taken together is the group wherein R is methyl and R is methyl, ethyl, phenyl, or when taken together with R and the carbon atom to which they are attached, cyclohexyl.

3. Compounds according to claim 2 wherein OR and R taken together is the group wherein R is methyl and R is methyl.

4. Compounds according to claim 2 wherein OR and R taken together is the group wherein R is methyl and R is ethyl.

5. Compounds according to claim 2 wherein OR and R taken together is the group wherein R is methyl and R is phenyl.

6. Compounds according to claim 2 wherein OR and R taken together is the group wherein R and R taken together with the carbon atom to which they are attached, is cyclohexyl.

7. Compounds according to claim 1 wherein R is hydrocarbon carboxylic acyl of less than 12 carbon atoms and R is tar-hydrocarbon carboxylic acyloxy of less than 12 carbon atoms.

8. Compounds according to claim 7 wherein R is acetyl and R is acetoXy.

9. A -pregnadiene-3fi,16a,17ot-triol-2O-one.

10. 6 halo A -pregnadiene-3fi,16a,17a-trio1-20-one wherein the halogen has an atomic number less than 53.

11. 6-chloro-A -pregnadiene-3B,16a,17or-triol-20-one.

12. 6 chloro-A -pregnadiene-3p,16a,l7u-triol-20-one 3-acetate.

13. 6 chloro 16a,17wisopropylidenedioxy-A -pregnadien-3/3-ol-20-one.

14. 6 chloro 16a,17a-is0propylidenedioxy-A -pregnadien-3/i-ol-20-one 3-acetate.

References Cited UNITED STATES PATENTS 3,314,944 4/1967 Diassi et al 260397.4 3,326,902 6/1967 Diassi 260397.4 3,126,399 3/1964 Sollrnan 260397.4 3,139,424 6/1964 Zderic et a1. 260397.4 3,198,792 8/1965 Reerink et al. 260239.5

LEWIS GOTTS, Primary Examiner.

J. R. BROWN, Assistant Examiner. 

